Measuring dynamic hormone release from pancreatic islets using perifusion assay
An important aspect of studying pancreatic islet physiology is to be able to measure hormone release from islet cells in response to specific stimuli. We demonstrate here step-by-step how to quantitatively measure hormone release from viable isolated pancreatic islets using the in vitro perifusion assay. Islet perifusion has two great advantages over other existing assays: (1) It allows pharmacological manipulation to dissect out signaling mechanisms underlying release of different hormones in the pancreatic islets and, (2) it allows performing simultaneous experiments on multiple islet preparations resulting in high throughput readouts. Other practical applications of islet perifusion include assessment of islet function and viability for experimental purposes and/or clinical application in human islet transplantation. The experimental conditions during islet perifusion are adjusted to simulate the islet “physiological” environment within the pancreas. Therefore, the overall time required to execute the described protocol will be more than 1 h to establish such conditions but should not exceed 2 h to avoid compromising islet health.
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Posted 10 Oct, 2011
Measuring dynamic hormone release from pancreatic islets using perifusion assay
Posted 10 Oct, 2011
An important aspect of studying pancreatic islet physiology is to be able to measure hormone release from islet cells in response to specific stimuli. We demonstrate here step-by-step how to quantitatively measure hormone release from viable isolated pancreatic islets using the in vitro perifusion assay. Islet perifusion has two great advantages over other existing assays: (1) It allows pharmacological manipulation to dissect out signaling mechanisms underlying release of different hormones in the pancreatic islets and, (2) it allows performing simultaneous experiments on multiple islet preparations resulting in high throughput readouts. Other practical applications of islet perifusion include assessment of islet function and viability for experimental purposes and/or clinical application in human islet transplantation. The experimental conditions during islet perifusion are adjusted to simulate the islet “physiological” environment within the pancreas. Therefore, the overall time required to execute the described protocol will be more than 1 h to establish such conditions but should not exceed 2 h to avoid compromising islet health.
Figure 1
Figure 2
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